Optimization of Electroporation Parameters for Cell Culture in Suspension

Objective To optimize the electroporation parameters in mouse primary lymphocyte cultured in suspension with adenosine deaminase acting on RNA 1(ADAR1) as a reporter gene. Methods Specific siRNA targeting ADAR1 was transfected into mouse primary lymphocyte by electroporation under the different experimental conditions including the voltage,the temperature and the state of cell vitality. The transfection efficiencies were evaluated under the reverse microscope,by cell counting and typan blue staining cell viability assay. Results The highest electroporation transfection rate was achieved under the conditions of voltage 400 V,electric time 40 μs for the mouse lymphocyte. The tansfection efficiencies were closely related to the vigorous state of the cells. The colder operation temperature could help better transfection efficiencies. Conclusion Electroporation is a highly efficient gene transfection method. Optimized electroporation parameters can improve the transfection efficiency.